Gene targeting is a genetic technique that utilizes homologous recombination between an engineered exogenous DNA fragments with the endogenous genome of an organism. In domestic animal, gene targeting has provided an important tool for producing Knock-out pig for GGTA1 gene to use xenotransplantation. The frequency of homologous recombination is a critical parameter for the success of gene targeting. The efficiency of homologous recombination in somatic cells is lower than that in mouse ES cells. So the application of gene targeting in somatic cells has been limited by its low efficiency. Recently, knock-out rat and mouse was generated by introducing nonhomologous end joining (NHE)-mediated deletion or insertion at the target site using zinc-finger nucleases (ZFN). Therefore, the development of effective knock-out and knock-in techniques in domestic animal is very important in biomedical research. In this present study, we investigated whether homologous recombination events occurs at cytidine monophospho-N-acetylneuraminic acid hydroxylase (CMAH) gene locus using ZFN in porcine primary fibroblast. CMAH-targeted ZFN DNA and mRNA were purchased from SIGMA-Aldrich. CMAH neo targeting vector consists of the neomycin resistance gene as a positive selectable marker gene, 789 bp 5’ arm and 763 bp 3’ arm from Exon 8 of CMAH gene. For transfection, the targeting vector and ZFN DNA or mRNA were introduced into ear fibroblasts cells of Chicago miniature pig by electroporation. After selection of G-418, PCR analysis was performed using 213 colonies transfected with ZFN DNA or mRNA. As a result, 39 positive colonies were identified in colonies transfected with ZFN DNA or mRNA. To our knowledge, this study provides the first evidence that the efficiency of gene targeting using ZFN was higher than that of conventional gene targeting in the porcine fibroblast. These cell lines may be used in the production of CMAH knock-out for xenotransplantation.

The purpose of this study was to determine the effects of heat appli˗ cation on the immune activities of the human body. To exam, further˗ more, the immune effect from the healthy volunteer(male:15, female:15) by monitoring changes of immune substances such as various leukocytes[total white blood cell(WBC), eosinophil, neutrophil, basophil, monocyte, and lymphocyte], a comparative study with warm water immersion(40.8±0.3℃) and infrared(250W) was carried out. The plasma analysis showed that the count of white blood cell, eosinophil, and neutrophil were elevated in warm water immersion- or infrared˗ stimulated group compared with control group. However, the count of basophil was decreased in both warm water immersion- and infrared-stimulated group than control group. Therefore, these results suggest that the thermostimulation improved immune activity.

본 연구는 돼지 B-casein 유전자 위치에서 EGFP가 발현될 수 있는 knock-in 벡터를 구축하기 위하여 실시되었다. 돼지의 B-casein 유전자를 이용하여 knock-in 벡터를 구축하기 위해 돼지의 태아 섬유아세포로부터 B-casein 유전자를 동정하였고 EGFP, SV4O polyA signal을 동정하였다. Knock-in 벡터는 5' 상동 영역 약 5 kb와 3' 상동 영역 약 2.7 kb로 구성되어있으며, positive selection marker로 neor 유전자를, negative selection marker로 DT-A 유전자를 사용하였다. 구축된 knock-in 벡터로부터 EGFP의 발현을 확인하기 위하여 생쥐 유선 세포인 HC11 세포에 knock-in 벡터를 도입하였다. 그 결과 EGFP의 발현을 HC11 세포에서 확인하였다. 이와 같은 결과로서 이 block-in 벡터는 knock-in 형질전환 돼지를 생산하는데 사용될 수 있을 것으로 생각된다.

본 연구는 사람 H-transferase가 과발현하는 돼지 체세포주를 개발하는데 있다. 돼지 세포에 사람 H-transferase 유전자를 발현시키는 것은 이종간 장기 이식에 있어서 초급성 거부 반응을 방지하기 위한 한 가지 방법이다. 본 연구에서는 과발현 벡터를 구축하기 위하여 사람 H-transferase을 HepG2 세포로부터 동정하였으며, 이 유전자를 CMV promoter를 이용하여 발현할 수 있도록 포유동물 발현 벡터인 pRc/CMV 벡터에 삽입하였다. 또한, 돼지 산자의 귀 세포를 이용하여 체세포를 수립한 후 jetPEI DNA transfection reagent를 이용하여 벡터를 도입하였고, 300 μg/ml의 G418로 12일간 선별하였다. PCR을 이용하여 선별된 colony들을 분석한 결과, 벡터가 도입되었음을 확인하였고, RT-PCR을 이용하여 사람 H-transferase mRNA가 발현하는 것을 확인하였다. 본 연구에서 확립된 세포주는 사람 H-transferase가 과발현하는 형질 전환 돼지의 생산에 이용될 수 있을 것으로 생각된다.

뿔나방과의 Monochroa 속에 대해 분류학적으로 정리한 결과 5종의 미기록종(M. cleodoroides, M. divisella, M. japonica, M.subcostipunctella and M. suffusella)을 포함한 6종의 국내 분포종을 확인, 정리하였다. 각종의 분류동정을 위한 검색표와 함께 암.수 생식기를 도해하였다.

This study was conducted to apply a horticultural therapy program to childhood cancer patients, to analyze their psychosocial changes and thus to examine its possibility as a way to manage the recovery of childhood cancer patients. This study was conducted on patients who were diagnosed with childhood cancer, and they were divided into the experimental group (8 patients who visited H association offices located in 3 cities and participated in the horticultural program) and the control group (5 patients who did not participate in the program). The homogeneity between the two groups before conducting the program was compared, and there was no statistically significant difference, which proved that the two groups were homogeneous. After conducting the program, the score of social immaturity, one of the sub-categories of problem behaviors, of the experimental group was statistically significantly lower than that of the control group (p=.019). Changes before and after the program within the groups were compared, and the somatization score (p=.039) of the experimental group (p=.039) after the program was 1.13, down from 2.13 before the program. The social immaturity score (p=.017) of the experimental group after the program was 1.00, down from 2.63 before the program, and the attention problem score (p=.026) of the group after the program was 1.25, down from 2.50 before the program. The total problem behavior score (p=.034) of the experimental group after the program was 9.63, down from 18.50 before the program, showing a statistically significant decrease. The cumulative subjective mood score (p=.000) of the experimental group throughout the entire sessions after the program was 7.82, up from 6.99 before the program, showing a statistically significant increase. Therefore, it can be concluded that the horticultural therapy program developed based on the ‘relaxationsocial support-cognitive behavior’ strategy was effective to reduce the negative psychosocial state of childhood cancer patients.

Light is very important for plant growth and a major limiting factor in indoor plant cultivation, such as in balconies. This study was conducted to investigate the light intensity and lettuce (Latuca sativa L.) growth in an apartment balcony affected by the direction of balcony. Photosynthetic photon flux (PPF), air temperature, and relative humidity in the east-, south-, or west-facing balcony were measured for a year. The lettuce was grown in the east-, south-, or west-facing balcony. The time and amount of light entering into the balcony differed according to the direction of the balcony window. Whereas light entered into the east-facing balcony from sunrise to noon, it entered into the west-facing balcony afternoon before sunset. The temperature and humidity in the balcony also varied according to the direction of the balcony window. Daily Light Integral (DLI) in the south-facing balcony was 20~50% of outdoor and DLI in east- or west-facing balconies was 10~20%. Also, the environmental characteristics of the balconies were affected seasonally. In the spring (from March to May), the DLI was around 10 mol･m-2･day-1 and the average temperature was around 25°C. But the DLI was below 10 mol･m-2･day-1 and the average temperature and relative humidity were around 30°C and over 60%, respectively, in the summer (from June to August). In the balcony, the average temperature was maintained around 10°C even in winter, which is a condition of freezing outdoor. Growth of the lettuce in the south-facing balcony was better than that of the other directions. This is attributed to better environmental conditions of south-facing balcony. Therefore, considering the environmental characteristics of the balcony, selection and management of crops with relatively low light demand like leafy vegetables are required. It is also recommended to cultivate crops from autumn to spring.

This study was conducted to analyze the muscle activity in the right forearm and shoulderwhile doing the sowing activity by muscle type and work section, and to find the possibility of the utilization of sowing as a muscle exercise and rehabilitative activity based on the results of the analysis. The results showed that the activity of 7 muscles including the upper trapezius near the neck, the upper trapezius near the shoulder, the middle deltoid, the biceps brachii, the triceps brachii, the brachioradialis, and the flexor carpi ulnaris were significantly higher than the stable state. In addition, the muscle activity in the sections of mixing the soil, filling the tray with the soil, sowing, writing the label, and watering was significantly higher than the stable state. In particular, the muscle activity of the biceps brachii and the triceps brachii was statistically significantly high in the section of filling the tray with the soil, which indicates that the task of filling the tray with the soil can be utilized as an exercise to activate the biceps brachii and the triceps brachii. In addition, the muscle activity of the brachioradialis was significantly high in all the work sections, which indicates that the sowing process can be utilized to exercise the brachioradialis in particular. These results can be used as the basic data about the activated muscle types and degrees throughout the sowing process. These results can be also used to induce the activation of specific muscles, and the overall exercise and rehabilitation of the forearms.